The kidney adjusts K+ secretion in the distal nephron in response to variations in dietary intake to maintain K+ homeostasis. The distal K+ secretion involves K+ efflux from the cell into the lumen through apical K+ channels such as ROMK. Na+ reabsorption via the epithelial Na+ channel ENaC provides the electrical driving force for K+ secretion. An increase in the activity of the Na-Cl cotransporter NCC may decrease Na+ delivery to ENaC thus diminishing K+ secretion. In support for the role of NCC as well as ROMK in maintaining K+ homeostasis during dietary variations of K+ intake, high K+ loading in rodents increases the density of ROMK while decreasing the density of NCC in the distal nephron. The upstream regulation of ROMK and NCC remains incompletely understood. WNK1 is a protein kinase of which gene mutations resulting in increased expression cause pseudohypoaldosteronism type II (PHA2), an autosomal-dominant disease characterized by hypertension and hyperkalemia. WNK1 has several alternatively spliced variants including a ubiquitous full- length long WNK1 (L-WNK1) and a shorter kidney-specific WNK1 (KS-WNK1). Cell-based expression studies have shown that L-WNK1 activates NCC and inhibits ROMK. KS-WNK1, by itself, does not regulate NCC and ROMK but reverses L-WNK1-mediated activation and inhibition of NCC and ROMK, respectively. The long- term goal of our research is to understand K+ homeostasis in the normal physiology and in diseased states. Here, we will examine three specific aims. Aim 1 is to examine the physiological role and mechanism of L- WNK1 in the regulation of renal Na+ and K+ transport. Aim 2 is to examine the hypothesis that KS-WNK1 antagonizes L-WNK1 regulation of renal Na+ and K+ transport and changes in dietary K+ affect the ratio of L- over KS-WNK1 to regulate NCC and ROMK. Aim 3 is to examine the interplay between aldosterone and L- and KS-WNK1 pathway in the regulation of renal Na+ and K+ transport and the response to variations in dietary K+. We have generated mouse models with genetically altered expression of L-and/or KS-WNK1. Renal Na+ and K+ transport in these mice will be studied by measuring blood and urinary electrolytes and expression of Na+ and K+ transporters in response to dietary Na+ or K+ perturbations and to diuretics. K+ secretion will also be studied by in vitro microperfusion of isolated CCD tubules. Surgical adrenalectomy will be performed to examine interactions between aldosterone and WNK1 pathway. These studies will provide important insights to the in vivo role of WNK1 in renal Na+ and K+ transport in the normal physiology and in diseased states. PUBLIC HEALTH RELEVANCE: The kidney plays an important role in regulating blood K levels by controlling urinary excretion. The goal of our research is to understand how the kidney regulates K excretion in the normal physiology and in diseased states. Our research may lead to better understanding of the kidney, heart and muscle diseases and potential new therapy for these diseases.